Characterization of the BPI-like gene from a subtracted cDNA library of large yellow croaker (Pseudosciaena crocea) and induced expression by formalin-inactivated Vibrio alginolyticus and Nocardia seriolae vaccine challenges.
One expressed sequence tag (EST 64LF004 clone), which is from the subtracted cDNA library of the head kidney of large yellow croaker (Pseudosciaena crocea) stimulated with peptidoglycan (PG) by suppression subtractive hybridization (SSH) method, was cloned using RACE-PCR. The full length cDNA, which possesses typical structural features of a signal peptide, a conserved LPS binding domain and two bactericidal permeability-increasing (BPI) motifs as in higher vertebrates, was identified as a novel homologue, namely of the large yellow croaker BPI-like molecule (Pc-BPI-L). Phylogenetic analysis showed this Pc-BPI-L of large yellow croaker as the most ancestral branch in bony fish clade. The recombinant Pc-BPI-L protein expressed in the Tn-5B1-4 insect cells was successfully produced and confirmed to have the predicted size of 52 kDa by Western blot analysis. At the message level, Pc-BPI-L mRNA was ubiquitously expressed in all tissues examined. Following formalin-inactivated Vibrio alginolyticus and Nocardia seriolae treatment, Pc-BPI-L message was differentially up-regulated in primary immune organs. These results indicate that Pc-BPI-L might be involved in the immune response to bacterial infection.
Huang
Yanqing
Y
Laboratory of Marine Life Science and Technology, College of Animal Sciences, Zhejiang University, 268 Kaixuan Road, Hangzhou 310029, PR China.
Lou
Huifang
H
Wu
Xinzhong
X
Chen
Yanxia
Y
D000595:Amino Acid Sequence; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000235:genetics* / Q000276:immunology; D001428:Bacterial Vaccines / Q000276:immunology* / Q000494:pharmacology; D001483:Base Sequence; D001798:Blood Proteins / Q000235:genetics* / Q000276:immunology; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D008969:Molecular Sequence Data; D009615:Nocardia / Q000276:immunology*; D009617:Nocardia Infections / Q000276:immunology / Q000382:microbiology / Q000662:veterinary*; D010473:Perciformes / Q000235:genetics* / Q000276:immunology / Q000382:microbiology; D010802:Phylogeny; D012313:RNA / Q000737:chemistry / Q000235:genetics; D011994:Recombinant Proteins / Q000276:immunology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D016415:Sequence Alignment; D015164:Vaccines, Inactivated / Q000276:immunology / Q000494:pharmacology; D014735:Vibrio Infections / Q000276:immunology / Q000382:microbiology / Q000662:veterinary*; D044149:Vibrio alginolyticus / Q000276:immunology*
D023181:Antimicrobial Cationic Peptides; D001428:Bacterial Vaccines; D001798:Blood Proteins; D018076:DNA, Complementary; D011994:Recombinant Proteins; D015164:Vaccines, Inactivated; C021623:bactericidal permeability increasing protein; D012313:RNA